7-(1,4-cyclohexadienyl-acylamino)cephalosporins

ABSTRACT

THIS INVENTION RELATES TO NEW CEPHALOSPORIN DERIVATIVES AND METHODS FOR PREPARING THE SAME, SAID DERIVATIVES HAVING THE STRUCTURE   1-(R-CH=N-CH(-CO-Y)-)CYCLOHEXA-1,4-DIENE AND   1-(R-CH2-NH-CH(-CO-Y)-)CYCLOHEXA-1,4-DIENE   WHEREIN Y IS 7-ACA OR 7-ADCA, OR AN ESTER OR SALT THEREOF, AND R IS PHENYL, SUBSTITUTED PHENYL, NAPHTHYL, SUBSTITUTED ANPHTHYL, LOWER ALKYL OR CYCLOAKYL. THESE COMPOUNDS ARE USEFUL AS ANTIBACTERIAL AGENTS.

3,812,110 7-(l,4-CYCLOHEXADIENYL-ACYLAMINO) CEPHALOSPORINS Bong Kuk Lee,Old Bridge, N.J., Dewey D. Y. Ryu, Holliston, Mass., and Gennaro JohnMiraglia, East Brunswick, Harold I. Basch, Somerset, and Barid B.Mukherjee, East Windsor, N.J., assignors to E. R.

Squibb & Sons, Inc., Princeton, NJ.

No Drawing. Continuation-impart of abandoned application Ser. No.229,170, Feb. 24, 1972. This application Aug. 21, 1972, Ser. No. 282,096

Int. Cl. C07d 99/24 US. Cl. 260-240 G 8 Claims ABSTRACT OF THEDISCLOSURE This invention relates to new eephalosporin deriva- Thisapplication is a continuation-in-part of our co: pending applicationSer. No. 229,170, filed Feb. 24, 1972, now abandoned.

This invention relates to new cephalosporin derivatives of the formula Io S and g s @fw o=' -N omooocm wherein Z is hydrogen, lower alkyl or asalt forming ion, e.g., an alkali metal such as sodium or potassium, analkaline earth metal such as calcium or magnesium, or that of an organicbase such as dibenzylamine, N,N- dibenzylethylenediamine, or the like,and Q is or --NHCH R wherein R is aryl, substituted aryl, lower alkyl orcycloalkyl.

Thus, the cephalosporin derivatives of the invention include thefollowing sub-genuses:

0 0 O-CE-ii-Y and Q-CH-PJ-Y It I'm I 8H CH: I l 1',

III IV Patented May 21., 1974 Ice wherein Y is 7-aminocephalosporanicacid (7-ACA) or 7-aminodesacetoxycephalosporanic acid (7-ADCA) and R isphenyl, substituted phenyl, naphthyl, substituted naphthyl, lower alkylor cycloalkyl. The phenyl or naphthyl substituents include halogen,hydroxy, amido, alkanoyl of up to 7 carbon atoms, alkoxy of up to 7carbon atoms, or alkyl of up to 7 carbon atoms, preferably in the orthoor fi-position.

The lower alkyl groups represented by the above R groups includestraight or branched chain aliphatic hydrocarbon radicals having up toseven carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, amyl, hexyl, heptyl, and the like. The lower alkylgroups can include as substituents any of the aryl groups mentionedbelow as well as halogen (Cl, Br, I or F).

The term aryl includes phenyl or naphthyl. These radicals can include assubstituents at the ortho position halogen, hydroxy, alkanoic acid,lower alkoxy, amido or any of the alkyl groups mentioned hereinbefore.

For example, aryl radicals and substituted aryl radicals contemplatedherein include, but are not limited to, the following:

and

The term cycloalkyl includes monocyclic carbocyclic radicals having from3 to about 6 carbons such as cyclopropyl, cyclobutyl, cyclopentyl andcyclohexyl.

The compounds of formula III can be prepared by reacting a compound ofthe structure with an aldehyde of the structure VI RCH to form compoundsof the structure III In carrying out the above reaction, the reaction ofcompound V with the aldehyde VI is preferably carried out in an aqueousmedia such as in a mixture of water with a lower alkanol such asmethanol, ethanol or n-propanol, or ktones such as methylethyl ketone ormethyl isobutyl ketone. The reaction may be carried at temperaturesranging from about 0 to about 40 C. Compound V is employed in a ratio tocompound VI within the range of from about 1:1 to about 1:3, preferablyfrom about 1:1 to about 1:1.5 and optimally at about 1:1.1.

Compounds of formula HI can also be prepared by reacting a compound ofthe formula VII with an aldehyde of formula VI to form a compound of theformula VIII and thereafter reacting VHI with an acid halide compound ofthe structure xii 0 alkyl to form a mixed anhydride of the structure OOZO= ---N CHzO COCH.

OOZ

to form the formula III compounds.

The reaction of compound VII with the-aldehyde VI is carried out in thepresence of an aqueous-alcoholic solvent, such as a mixture of water andmethanol, at temperatures ranging from ambient temperature to theboiling point of the solvent. Compound VII can be employed in a molarratio to the aldehyde VI within the range of from about 1:1 to about 1:3and preferably from about 1:1 to about 121.5.

The reaction of compounds VIII and IX is carried out in a mixture ofsolvents such as acetone, dioxane and lutidine at temperatures rangingfrom about --10 to about 20 C. and preferably from about -10 to about 10C., employing a molar ratio of IX:VIII within the range of from about1:1 to about 3:1 preferably 1.1:1 to 1.5 :1.

The reaction of the mixed anhydride X with 7-ACA or 7-ADCA is carriedout in an aqueous solvent such as aqueous sodium bicarbonate at atemperature with the range of from about 15 to about C. and preferablyfrom about -5 to about 0 C. employing a molar ratio of XzXI or XzXIIwithin the range of from about 1:08 to about 2:1 and preferably fromabout 1.1:1 to about 1.5:1.

Compounds of formula III can be reduced by reacting III with a reducingagent such as sodium borohydride, aluminum borohydride, lithium aluminumhydride, or hydrogen in conjunction with a catalyst for reduction suchas platinum or palladium to form compounds of the structure The abovereduction can be carried out in water or aqueous solvents, such asaqueous potassium phosphate, at temperatures ranging from about 0 toabout 40 C.

and preferably from about 10 to about 20 C. employing-a-molar-ratio ofcompound III to reducing-agent within the range from about 1:2 to about1: 10 and preferably from about 1:4 to about 1:6.

Alternatively, the formula IV compounds can be prepared by formingcompounds of formula VIII above, reducing the formula VIH compound byreacting it with any of the aforementioned reducing agents to, form acompound of the formula XIII I r @a oz I'm an.

and thereafter reactingXIII with an acid halide of structure IX to formthe mixed anhydride of formula XIV.

XIV V '0 o @on -ii-o-b-oanwr d m l I which can be reacted with 7ACA or7-ADCA to form the formula IV compounds.

It will be appreciated that essentially the same reaction conditions asset out with respect to the reaction of compounds VIII, IX, X and XIapply here as well.

The starting materials-of structure V are disclosed in US. Pat. No.3,485,819 to Weisenborn, et al., and are prepared by coupling a compoundof the structure Xv Q-orb-cooz s'm, with a 7ACA or'7-ADCA moiety, thatis'with xr s I NH2- O=---N CH:

XII v s OOZ as described in Pat. No. 3,485,819.

Examples of aldehydes which can be employed herein as startingmaterialsinclude, but are not limited to, the following cgHscHo;

C(HQCHO CEO and

A-CHO The products of this invention form salts which are also part ofthe invention. Basic salts form with the acid moiety as discussed abovein connection with the symbol Z. Acid addition salts also form with thea-amino nitrogen. Such acid salts include, for example, inorganic saltssuch as the hydrohalides, e.g., hydrobromide, hydrochloride,hydroiodide, sulfate, nitrate, phosphate, borate, etc., and organicsalts such as acetate, oxalate, tartrate, malate, citrate, succinate,benzoate, ascorbate, methanesulfonate and the like. It is frequentlyconvenient to isolate and purify the product by forming a soluble orinsoluble salt, as desired, then regenerating the free compound, byneutralization, for example.

The compounds of this invention have a broad spectrum of antibacterialactivity against both gram positive and gram negative organisms such asStaphylococcus aureus, Salmonella schottm uelleri, Pseudomonasaeruginosa, Proteus vulgaris, Escherichia coli and Streptococcuspyogenes. They may be used as antibacterial agents in a prophylacticmanner, e.g., in cleaning or disinfecting compositions, or otherwise tocombat infections due to organisms such as those named above, and ingeneral may be utilized in a manner similar to penicillin G and otherpenicillins and cephalosporins. For example, a compound of Formula I ora physiologically acceptable salt thereof may be used in various animalspecies in an amount of about 0.1 to 100 ing/kg. daily, orally orparenterally, in single or two to four divided doses to treat infectionsof bacterial origin. -By way of illustration the P-D orally in mice in asingle administration is 1.3 mg./ kg. against Streptococcus, 8.6mgL/k'g. against Proteus and 11.8 mg./ kg. against Salmonella,respectively. Up to about 600 mg. of a compound of Formula -I or a saltthereof may be incorporated in an oral dosage form such as tablets,capsules or elixirs or in :an injectable form in a sterile aqueousvehicle prepared according to conventional pharmaceutical practice. Incleaning or disinfecting compositions, e.g., in barns or dairyequipment, a concentration of about 0.01 to 1% by weight of suchcompounds admixed with, suspended or dissolved in conventional inert dryor aqueous carriers for application by washing or spraying may be used.

The following examples are illustrative of the invention. Alltemperatures are on the centigrade scale.

6 EXAMPLE 1 7 [2 (1,4 cyclohexadien 1-yl)-2-[[(2-hydroxy1-naphthyl)methylene]amino]acetamido] 3 methyl 8 oxo 5 thial-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, sodium salt, hydrate (A)Na salt of a-amino-2,5-dihydrobenzyl-3-desacetoxycephalosporin Thesodium salt of a-amino-2,5-dihydrobenzyl-3-desacetoxycephalosporin(sephradine) is formed by dissolving 1396 mg. (4 millimoles) ofa-amino-2,5-dihydrobenzyl-3-des'acetoxycephalosporin and 336 mg. (4millimoles) of NaI-ICO in a mixture of 30 ml. H 0 and 350 ml. methanol.

(B) 7 [2 1,4 cyclohexadien 1 yl)-[[2-hydroxyl naphthyl)methylene]amino]acetamido] 3-methyl- 8 oxo 5 thio 1-azabicyclo[4.2.0]oct-2-ene-2-can boxylic acid, sodium salt, hydrate EXAMPLE 2 7[2 (1,4 cyclohexadien l-yl)-2-[[(2-hydroxy-1-naphyl)methyl]amino]acetamido] 3 methyl-8-oxo 5 thia 1azabicyclo[4,2.0]oct. 2-ene-2-carboxylic acid, sodium salt, hydrate1,019 mg. (1.83 millimoles) of the product of Example 1 is dissolved in400 ml. of potassium phosphate buffer (0.5 M, pH 6), and to thissolution is added, 325 mg. (8.55 millimoles) of NaBH dissolved in 15 ml.H O, dropwise, for 3 hours, with stirring, at a temperature of about 10C. The reaction mixture is acidified to pH 3 in an ice bath, andcentrifuged to separate a light brownish solid, 6.70 mg. (71% yield) ofsolid is isolated. After washing the solid twice with 5 ml.-portions ofcold water, it is dried in vacuo. The resulting solid, 432.3 mg. (0.838millimole), and 73.2 mg. (0.872 millimole) of NaHCO are dissolved in amixture of 10 ml. H 0 and 30 ml. methanol. The methanol is removed invacuo at a temperature below 10 C. The remaining solid, weighting 380mg. (88% yield) is obtained, which analyzes as follows: (C H N SO NaHO): Calcd.: 27 C, 59.44; 26 H, 5.17; 3 N, 7.70; 1 S, 5.82; 1 Na, 4.22.Found: 27 C, 59.14; 26 H, 5.32; 3 N, 8.01; 1 8,603; 1 Na, 3.69.

EXAMPLE 3 7 [2 1,4 cyclohexadien1-yl)-2-[[(2-hydroxy-lnaphthyl)methyleneJamino] acetamido] 3 methyl-8-oxo 5 thia 1 azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, sodium salt,hydrate A suspension of the sodium salt of D-2-amino-2-(l,4-cyclohexadienyl)-acetic acid 1.75 g. (10 millimoles) andZ-hydroxy-l-naphthaldehyde 3.44 g. (20 millimoles) in a mixture ofethanol (250 ml.) and methanol (20 ml.) is refluxed for 2 hours. Themixture is then evaporated under reduced pressure and the residue iswashed with ether and recrystallized from ethanol to give crystals ofthe sodium salt of (1,4-cyclohexadien-1-yl) [[(Z-hydroxy-1-naphthyl)methylene]amino1acetic acid 3.11 g. yield).

This N-protected amino-acid 2.63 g. (8 millimoles) is dissolved in amixture of dry acetone ml.), dioxane (25 ml.) and 2,6-lutidine (1.35ml.) The solution is rapidly chilled to 5, treated with ethylchlorocarbonate 0.955 mg. (0.84 ml., 8.8 millimoles) and stirred at 0for 10 minutes, during which lutidine hydrochloride 7 is precipitatedand the mixed anhydride formed in solution, The suspension is thencooled-to -45 and stirred 'vigorouslywhile an ice-cold solution of.7-ADCA 1.31 g.

.ether extracts, containing the N-protected cephalosporin are washedwith water (10 mli) and'then extractedwith suflicient 3% sodiumbicarbonate to give a neutral aqueous phase, which is separated andevaporated at low temperature and pressure. The residue 1.71 g. (3.16millimoles, 40% yield) is shown,'by paper chromatography of a smallportion, to contain the sodium salt-of the cephalosporin of the abovetitle.

I EXAMPLES 4 TO 12 The procedure described in Example 1 is repeated withthe exception that the aldehydes set out below in Table I,

column A areemployed, to thereby form the products set out in column BofTableI.

t N a.

a H it 002 TABLE I Column A Column B Aldehyde(R-CHO) Product Examplenumber R R Y 4 AS in column A.

5 CHO Do.

I Do.

I Do.

CsHzCHO D0.

CaHnCHO D0- A4110 12- (not. H9

8 EXAMPLES 13 21 Employing the procedure described in Example 2, butsubstituting as the st'arting'mater'ial, the products of Examples 4 to12, the products set out in Table II below are obtained.

( OONa TABLE 11- Example number R v EXAMPLES 'zjz 'ro so Employing theprocedure of Example 3, but substitut ing the starting material set outin column A of Table 9 III below, the product shown in column B thereofis obtained.

TABLE III 10 (B) The product from step A (600 mg., 1.83 mm.)

is suspended in a mixture of ml. dry acetone, 6 ml. p-

Starting material Product l CH-C-NHq N I f 0 N OH s I 11H R l R O 0 ZExample number R Z R Z 22 33 K As per columnA 23 3 H As per column A 24CH| As per column A 25 33 Na As per column'A 26 H As per column A -C;H1CaH As per column A -C;H Na As per column A K As per column A 30 Na Asper column A EXAMPLE 31 dioxane, and 0.35 ml. 2,6-lutidine. Thesuspension is N-[2-(l,4-cyclohexadien 1 yl) 2 [[(2-hydroxy-1-naphthyl)rnethylene]amino]acetyl] 7 aminocephalosporanic acid cooled to5, 0.25 ml. of ethylchloroformate are added and the suspension stirredfor 10 minutes at 0. 7ACA (472 mg., 1.83 mm.) dissolved in 20 m1. of a1% NaHCO3 l. T EH 0=N CHQOCOCH,

O O Nat-2H2 0 thyl) methyl]amino]acetyl].;7-amin0 ;phalospor anic acidThe product from part B of Example 31 (152 mg., .252 mm.) is dissolvedin 10 ml. of potassium phosphate buffer (0.05 M, pH 6) and placed in anice bath. NaBH (28.7 mg., 0.756 mm.) dissolved in 2.5 ml. H Ois added,dropwise, with vigorous stirring for 1 hour. The mixture is thenacidified to pH 2 with 1 N HCl and filtered. The resulting solid iswashed 3 times using 10 ml; portions of cold water for each wash, andonce with 10 ml. of-cyclohexane. The product is a brown solidweighing 72mg., M.P. 197202 having the formula EXAMPLES 3 3--4 l The procedure ofExample 31 is repeated except substituting for2-hydroxy-l-naphthaldehyde the respective aldehydes of Examples 4-12.The product 'in each case is the 7-ACA analog of the 7-ADCA product ofExamples 4-12.

EXAMPLES 42-50 R is alkyl of from 1 to 7 carbon atoms, cycloalkyl offrom 3 to 8 carbon atoms or 2-hydroxynaphthyl, R is cycloalkyl of from 3to 8 carbon atoms, phenyl, naphthyl, or substituted phenyl orsubstituted naphthyl wherein the substituent is F, Cl, Br, I, hydroxy,carbamyl, alkanoyl of up to 7 carbon atoms, alkoxy of up to 7 carbonatoms or alkyl of up to 7 carbon atoms, or substituted alkyl wherein thesubstituent is F, Cl, Br, I, hydroxy, or carbamyl, W is CH or CH OCOCHand Z is hydrogen, alkyl of from 1 to carbon atoms, an alkali oralkaline earth metal, dibenzylamine or N,N-dibenzylethylenediamine.

2. A compound according to claim 1 of the formula wherein R, W and Z areas defined in claim 1.

3. A compound according to claim 1 of the formula wherein R, W and Z areas defined in claim 1.

4. A compound according to claim 2 of the formula 5. A compoundaccording to claim 3 of the formula I S 1.. l O= N CHzOCOCH:

(BOOZ 6. A compound according to claim 2 of the formula S Ge N O I OH OC OCH (ll/ H --N 2 8 7. A compound according to claim 3 of the formula8. A compound of claim 3 wherein R is o-substituted HENRY R. JILES,Primary Examiner S. D. WINTERS, Assistant Examiner r l GGllBa "H050,UNITED STATES PATENT FICE CERTIFICATE OF CORRECTION Patent No. 3312, 110Dated 1 May 21, I 1974 Inventor(s) B g k et al.

It is certified that error appears in the above-identified patent andthat said Letters Patent are hereby corrected as shown below:

F Column 1, lines 20-25, the formulas should read P I-I.C-Y and Column1, lines 65-70, the formulas should read Signed and sealed this 29th dayof October 1974.

(SEAL) Attest:

C- MARSHALL DANN Commissioner of Patents McCOY M. GIBSON- JR. AttestingOfficer

